Esters for treatment of ocular inflammatory conditions

ABSTRACT

The present invention relates to ophthalmic compositions and methods for the treatment of dry eye and other inflammatory ocular conditions. In particular, the present invention relates to a composition comprising an esterified anti-inflammatory lipid mediator, which is an ester of an anti-inflammatory lipid mediator that is a reaction product of the anti-inflammatory lipid mediator and a monohydric alcohol or an amide wherein the majority of the anti-inflammatory lipid mediator is present in an ester form. In this way, the compositions are substantially free of an acid form of the anti-inflammatory lipid mediators. This composition can be topically delivered to the ocular surface via a preparation, solution, gel, ointment, and/or strip and/or a contact lens.

RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.13/495,049, filed on Jun. 13, 2012, which claims priority to U.S.Provisional Patent Application Ser. No. 61/503,158, filed on Jun. 30,2011.

TECHNICAL FIELD

The present invention relates to ocular products containing esterifiedanti-inflammatory lipid mediators for relief of dry eye or treatment ofinflammatory ocular conditions. Provided is an ophthalmic compositionthat comprises an esterified anti-inflammatory lipid mediator. Furtherprovided are sterile preparations, solutions, gels, ointments, and/orstrips for administration to the eye and/or a contact lens thatcomprises an esterified anti-inflammatory lipid mediator.

BACKGROUND

It is known that sufficient lubrication is necessary for good eyehealth. Tears nourish the ocular tissues and protect the surface of theeye from foreign bodies. Changes in the ocular surface due toalterations in the quality or quantity of tears (caused by eitherdecreased tear production or increased tear film evaporation) can leadto dry eye syndrome and other inflammatory ocular conditions. Typicalsymptoms of dry eye and other inflammatory ocular conditions includedryness, burning, itchiness, scratchiness, stinging, a sandy/grittysensation, tired eyes, and sensitivity to light. These symptomstypically worsen as the day progresses. Other symptoms include pain,redness, a pulling sensation, pressure behind the eye, and a feelingthat there is something in the eye. Because of the range of symptoms,individuals suffering from dry eye and other inflammatory ocularconditions often complain of eye irritation and discomfort.

If dry eye and other inflammatory ocular conditions are left untreated,it can produce complications that can cause eye damage, resulting inimpaired vision or (rarely) the loss of vision. When symptoms aresevere, they can interfere with the quality of life of an individualsuffering from dry eye.

The ocular surface is normally covered by a tear film—the clear liquidthat coats the outer tissues of the eye. The tear film is composed ofthree layers; the most superficial layer of the tear film is the lipidlayer, which covers the aqueous layer of the tear film, and then thethird layer is a mucinous layer. Any abnormality in any one of the threelayers, particularly a disturbance in the lipid layer, produces anunstable tear film, which results in symptoms of dry eye and otherinflammatory ocular conditions.

Current methods of alleviating the symptoms of dry eye includeadministering artificial tears to the ocular surface. These artificialtears, however, must be administered every few hours, and only providetemporary and incomplete relief of the symptoms of dry eye. Thus, thereis a need for compositions and methods to treat various eye disordersand conditions, including but not limited to, dry eye syndrome and otherinflammatory ocular conditions.

It has been noted that consumption of dark fleshed fish containingdietary omega-three fatty acids is associated with a decreased incidenceof dry eye symptoms. Omega-three and omega-six fatty acids are compoundsknown as “essential” fatty acids because they are essential to humanhealth. These fatty acids, however, are not produced by the human body;instead, the fatty acids can be introduced into the body via dietaryintake, either in the form of food or as supplements. Oral consumptionof omega-three fatty acids, however, does produce potential side effectssuch as effects on bleeding time, increasing cholesterol (LDL) level,high caloric intake, a fishy aftertaste, and gastrointestinaldisturbances. Because of their potential to improve the symptoms of dryeye and other inflammatory ocular conditions, work on omega-three fattyacids when used in a topical application to the ocular surface has shownpromising results. (Rashid, S. et al., “Topical Omega-3 and Omega-6Fatty Acids for Treatment of Dry Eye,” Arch Opthalmol. 2008;126(2):219-225). Using topical formulations of fatty acids to treat dryeye would provide more flexibility for treatment, including lesseningside-effects that patients can experience from oral intake of fattyacids.

Omega-three fatty acid-containing oils such as botanical oils have beenused to form non-irritating ophthalmic compositions (e.g., U.S. PatentApplication Pub. 2010/0305045 (Abbott Medical Optics, Inc.)). Hydrogelcontact lenses can comprise a polymeric matrix and a hydrophobic comfortagent distributed in the polymeric matrix, where the hydrophobic comfortagent can include a monoglyceride, a diglyceride, a triglyceride, aglycolipid, a glyceroglycolipid, a sphingolipid, a sphingoglycolipid, aphospholipid, a fatty acid, a fatty alcohol, a hydrocarbon having aC₁₂-C₂₈ chain in length, a mineral oil, a silicone oil, or a mixturethereof. (U.S. Patent Application Pub. 2010/0140114 (Ciba VisionCorporation)). Ophthalmic lenses have been provided with anti-toxinagents that are monoesters and/or diesters of a polyhydric aliphaticalcohol and a fatty acid containing from eight to eighteen carbon atomsand wherein said monoester has at least one hydroxyl group associatedwith its aliphatic alcohol residue (U.S. Pat. No. 5,472,703). Resolvinsand protectins have been used to help treat pathologies associated withangiogenesis and ocular neovascularization, particularly associated withretinopathy of prematurity (e.g., U.S. Patent Application Pub.20100105773 (Children's Medical Center Corp.)). Lipoxins have also beenused to treat pathologies associated with ocular neovascularization(e.g., U.S. Patent Application Pub. 20100105772 (Serhan, et. al.)).

Accordingly, there remains a need in the art for improved ocularproducts that relieve/mediate symptoms of dry eye and other inflammatoryocular conditions.

SUMMARY

In one embodiment, the present invention provides an ophthalmiccomposition for treatment of ocular conditions, the compositioncomprising an ester or amide of an anti-inflammatory lipid mediator thatis a reaction product of the anti-inflammatory lipid mediator and amonohydric alcohol or an amide. Generally, the majority of theanti-inflammatory lipid mediator is present in an ester form. This is incontrast to anti-inflammatory lipid mediators being present in an acidform. In one or more embodiments, the composition is substantially freeof fatty acids. Such an esterified anti-inflammatory lipid mediator canbe dispersible and/or dissolvable or emulsifiable in an aqueous deliverysystem.

In another embodiment, the present invention provides a sterilepreparation, solution, gel, ointment, emulsion or strip foradministration to the eye or a contact lens comprising an esterifiedanti-inflammatory lipid mediator.

In a further embodiment, the present invention provides a method oftreating, preventing or mitigating inflammatory ocular conditions and/ordry-eye in an individual in need thereof which comprises delivering tosuch individual's ocular surface a therapeutically effective amount of acomposition comprising an anti-inflammatory lipid mediator.

These and other embodiments of the invention will become apparent fromthe following description of the presently preferred embodiments. Thedetailed description is merely illustrative of the invention and doesnot limit the scope of the invention, which is defined by the claims andequivalents thereof. Variations and modifications of the invention maybe effected without departing from the spirit and scope of the novelcontents of the disclosure.

DETAILED DESCRIPTION

Provided are processes of making and using ocular products containingesterified anti-inflammatory lipid mediators, wherein the majority ofthe anti-inflammatory lipid mediator is present in an ester form. It hasbeen discovered that the use of esterified anti-inflammatory lipidmediators, when the majority of the anti-inflammatory lipid mediator ispresent in the ester form, results in an ocular product that greatlyimproves initial comfort upon contact with or administration to theocular surface. Ocular products include, but are not limited to,preparations, solutions, gels, ointments, emulsions, strips, ophthalmicdevices, and the like any which can be administered to the ocularsurface, including the eye.

With respect to terms used in this disclosure, the following definitionsare provided.

Reference to “anti-inflammatory lipid mediator” includes those moleculesthat play a role (directly or indirectly) in the inhibition of cytokineproduction by epithelial cells or immune cells, in the inhibition ofreactive oxygen species (ROS) production by epithelial cells or immunecells, in the control and/or inhibition of recruitment of white bloodcells (reduction in leukocytes infiltration), and/or in the resolutionof inflammation (promotion of uptake of dead cells). Suitableanti-inflammatory lipid mediators are generally acid-based entitieswhose carboxylic groups of the hydrocarbon chain can be esterified. Themajority of the anti-inflammatory lipid mediator is present in the esterform. Anti-inflammatory lipid mediators can be reacted with hydroxylgroups of various entities as desired. The hydroxyl groups are deliveredby monohydric alcohols that can provide therapeutic benefits to the eye,including osmoprotection, in conjunction with the esterifiedanti-inflammatory lipid mediators.

As used herein, the term “about” refers to a range of +/−5% of thenumber that is being modified. For example, the phrase “about 10” wouldinclude both 9.5 and 10.5.

As used herein, the use of “a,” “an,” and “the” includes the singularand plural.

As used herein, the term “ophthalmic composition” refers to a compoundor mixture suitable for administration to the eye or ocular surface.Ocular compositions include preparations, solutions, gels, ointments,emulsions, strips and the like.

As used herein the term “sterile preparation” includes any compound ormixture for direct administration to any part of a mammalian body,including implantation, injection, administration as a drop, gel orwash, and the like, wherein the preparation is substantially free fromundesired foreign matter just prior to administration. Methods forinsuring sterility include aseptic packaging and sterilization byexposure to radiation, heat combinations thereof and the like.

As used herein, the term “individual” includes humans and vertebrates.

As used herein, the term “agent” includes any compound, composition, tobe tested for efficacy in the methods disclosed herein.

As used herein the term “ocular surface” includes the wet-surfaced andglandular epithelia of the cornea, conjunctiva, lacrimal gland,accessory lacrimal glands, nasolacrimal duct and meibomian gland, andtheir apical and basal matrices, puncta and adjacent or relatedstructures, including the eyelids linked as a functional system by bothcontinuity of epithelia, by innervation, and the endocrine and immunesystems.

As used herein, the term “contact lens” refers to a structure that canbe placed on the cornea of an individual's eye. The contact lens mayprovide corrective, cosmetic, therapeutic benefit, including woundhealing, delivery of drugs or neutraceuticals, diagnostic evaluation ormonitoring, or UV blocking and visible light or glare reduction, or acombination thereof. A contact lens can be of any appropriate materialknown in the art, and can be a soft lens, a hard lens, or a hybrid lens.

As used herein, the term “silicone hydrogel contact lens” refers to acontact lens formed from a polymer comprising silicone containing andhydrophilic repeating units.

As used herein, the term “hydrogel” or “hydrogel material” refers to ahydrated crosslinked polymeric system that contains water in anequilibrium state. Hydrogels generally contain at least about 15 wt %water, and in some embodiments at least about 20 wt % water atequilibrium.

Conventional hydrogels are prepared from monomeric mixturespredominantly containing hydrophilic monomers, such as 2-hydroxyethylmethacrylate (“HEMA”), N-vinyl pyrrolidone (“NVP”), or vinyl acetate.U.S. Pat. Nos. 4,495,313, 4,889,664, and 5,039459 disclose the formationof conventional hydrogels.

As used herein, the term “silicone hydrogel” refers to a hydrogelobtained by copolymerization of at least one silicone-containingmonomer, macromer, prepolymer, with at least one hydrophilic component.Examples of silicone hydrogels include balafilcon, acquafilcon,lotrafilcon, comfilcon, galyfilcon, senofilcon, narafilcon, falcon II 3,asmofilcon A, as well as silicone hydrogels as prepared in U.S. Pat. No.5,998,498, WO 03/22321, U.S. Pat. Nos. 6,087,415, 5,760,100, 5,776,999,5,789,461, 5,849,811, 5,965,631. U.S. Pat. No. 7,553,880, WO2008/061992, and U.S. 2010/048847. These patents, as well as all otherpatents disclosed in this paragraph, are hereby incorporated byreference in their entireties.

Hard contact lenses are made from polymers that include but are notlimited to polymers of poly(methyl)methacrylate, silicon acrylates,fluoroacrylates, fluoroethers, polyacetylenes, and polyimides, where thepreparation of representative examples may be found in JP 200010055, JP6123860 and U.S. Pat. No. 4,330,383. Intraocular lenses of the inventioncan be formed using known materials. For example, the lenses may be madefrom a rigid material including, without limitation, polymethylmethacrylate, polystyrene, polycarbonate, or the like, and combinationsthereof. Additionally, flexible materials may be used including, withoutlimitation, hydrogels, silicone materials, acrylic materials,fluorocarbon materials and the like, or combinations thereof. Typicalintraocular lenses are described in WO 0026698, WO 0022460, WO 9929750,WO 9927978, WO 0022459, and JP 2000107277. All of the referencesmentioned in this application are hereby incorporated by reference intheir entireties.

A therapeutically effective amount of an anti-inflammatory lipidmediator is an amount effective to produce a clinically recognizablefavorable change in the pathology of the disease or condition beingtreated. A therapeutically effective amount includes those effective totreat, reduce, alleviate, ameliorate, mitigate, eliminate or prevent oneor more symptoms of the ocular conditions sought to be treated or thecondition sought to be avoided or treated.

One of skill in the art would readily be able to determine what is atherapeutically effective amount or an effective amount.

As used herein, the term “inflammatory ocular condition” includes dryeye syndromes, which is also called keratoconjunctivitissicca (KCS). Dryeye is a multifactorial disease of the tears and ocular surface thatresults in symptoms of discomfort, visual disturbance, and tear filminstability with potential damage to the ocular surface. It isaccompanied by increased osmolarity of the tear film and inflammation ofthe ocular surface. Dry Eye Syndrome (DES) is defined as a disorder ofthe tear film, resulting from tear deficiency and/or excessive tearevaporation, causing damage to the ocular surface and causing symptomsof ocular discomfort. There are two main forms of dry eye syndrome: teardeficiency forms (including Sjögren's syndrome and non-Sjögren's teardeficient) and evaporative forms. The tear film normally covers thefront part of the eye, namely the cornea and the conjunctiva. The tearfilm is constantly exposed to multiple environmental factors, includingvariable temperature, airflow, and humidity, which may stimulate orretard its evaporation. In particular, a low humidity setting in thepresence of a significant airflow increases the tear evaporation rate,as is frequently reported by subjects in desiccating environments.Indeed, even people with a normal tear secretion rate may experience dryeye symptoms while exposed to dry environments, such as in airplanes anddry workplaces.

Dry eye can also be defined as a condition with a decrease or change inquality of tears irrespective of the presence or absence of corneal andconjunctival lesions. It includes dry eye conditions found inindividuals who have hypolacrimation, alacrima, xerophthalmia, anddiabetes, HIV/AIDS etc.; post-cataract surgery dry eye; allergicconjunctivitis-associated dry eye; dry-eye associated with prolongedcontact lens use; and age-related dry-eye syndrome. Dry eye can alsoinclude the conditions found in hypolacrimation individuals induced bylong time visual display terminal (VDT) operations, room dryness due toair-conditioning, and the like. An “inflammatory ocular condition” canalso refer to, but is not limited to: keratoconjunctivitissicca (KCS),age-related dry eye, Stevens-Johnson syndrome, Sjögren's syndrome,ocular cicatricalpemphigoid, blepharitis, corneal injury, infection,Riley-Day syndrome, congenital alacrima, nutritional disorders ordeficiencies (including vitamin), pharmacologic side effects, eyestress, glandular and tissue destruction, environmental exposure (e.g.smog, smoke, excessively dry air, airborne particulates), autoimmune andother immunodeficient disorders, and comatose individuals renderedunable to blink.

As used herein “contact lens related dry eye” (“CLRDE”) is a disordermarked by at least one objective clinical symptom and at least onesubjective symptom. Clinical symptoms are selected from (a) a tear filmbreak up time (“TFBUT”) of less than about 10 seconds in at least oneeye; (b) a fluorescein staining score ≥3 on a scale of 0-15 in at leastone eye; (c) a lissamine green staining score ≥3 on a scale of 0-18 inat least one eye; or (d) a tear meniscus grade of ‘abnormal’ in at leastone eye. Subjective symptoms are determined via patient feedback andinclude (a) ≥about 2 hour difference between average daily contact lenswear time and average daily comfortable contact lens wear time and (b) arating of frequent or constant feelings of dryness, burning, stinging ordiscomfort during lens wear. CLRDE sign includes both excessive tearevaporation and Non-Sjogren's aqueous tear deficiency. Excessive tearevaporation is a disorder marked by a TFBUT of about 10 seconds or lessin at least one eye or a TFBUT of 10 seconds or less in at least one eyeas well as conjunctival or corneal staining of about 3 or greater on theNEI scale. Non-Sjogren's aqueous tear deficiency tear meniscus is adisorder marked by a grade of ‘abnormal’ in at least one eye or a tearmeniscus grade of ‘abnormal’ in at least one eye as well as conjunctivalor corneal staining of 3 or greater on the NEI scale.

As used herein the term “adnexal inflammation” includes inflammation ofany area or part of the eye or ocular system, including but not limitedto the eyelids, the lacrimal glands and extraocular muscles.

As used herein, there term “osmoprotection” means to maintain anophthalmic osmolarity within a normal physiological range (preferably270-320 mOsm/kg, with an average of about 290 mOsm/kg) and/or protectepithelial tissue against the effects of hypertonic conditions, wherethe unit “mOsm/kg” is milli-osmole per kilogram. Osmoprotectants, agentsthat offer osmoprotection, are generally uncharged, can be held withinan ocular cell, are of relatively small molecular weight, and areotherwise compatible with cell metabolism. Osmoprotectants protectagainst hypertonicity below the ocular surface and provide hydration tothe epithelial surface. Osmoprotectants include, without limitation,glycerol, inositol, sorbitol, xylitol, and erythritol.

As used herein, the term “unsaturated fatty acid” refers to a fatty acidcontaining at least one double or triple bond. Fatty acids in this classuse the Greek alphabet to identify the location of double bonds. The“alpha” carbon is the carbon closest to the carboxyl group and the“omega” carbon is the last carbon of the chain. For example, linoleicacid, and gamma-linolenic acid (LA and GLA respectively) are omega-sixfatty acids, because they have double bonds six carbons away from theomega carbon. Alpha-linolenic acid is an omega-three fatty acid becauseit has a double bond three carbon atoms from the omega carbon.

As used herein, the term “omega-three fatty acid” refers to fatty acidsthat have double bonds three carbon atoms from their omega carbon atom.For example, an omega-three fatty acid includes, but is not limited toalpha linolenic acid (ALA). Other omega-three fatty acids includederivatives of ALA. A “derivative” of ALA is a fatty acid that is madeby a chemical modification performed upon alpha linolenic acid by, forexample, an enzyme or is done by organic synthesis. Examples ofomega-three fatty acids that are derivatives of ALA, include but are notlimited to, eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), andthe like. An “omega-three fatty acid” can comprise one or moreomega-three fatty acids.

As used herein, the term “omega-six fatty acid” refers to one or morefatty acids that have a double bond 6 carbon atoms from their omegacarbon atoms. For example, an omega-six fatty acid includes, but is notlimited to linoleic acid (LA). Other omega-six fatty acids includederivatives of linolenic acid. A “derivative” of linoleic acid is afatty acid that is made by a chemical modification performed uponlinoleic acid. Examples of omega-six fatty acids that are derivatives oflinoleic acid include, but are not limited to, gamma linolenic acid(GLA), dihomogammalinolenic acid (DGLA), and the like. In someembodiments, the composition comprises at least one non-inflammatoryomega-six fatty acid. A non-inflammatory omega-six fatty acid is anomega-six fatty acid that does not promote or cause inflammation. Insome embodiments the inflammation is in the eye or affects the ocularsurface. One of skill in the art can determine if a fatty acid causes orpromotes inflammation. If the fatty acid causes or promotesinflammation, the fatty acid can be excluded from the composition.

As used herein the term “linoleic acid” refers to 9,12-octadecandienoicacid, which has a short hand designation of 18:2(n-6), which is numberof carbons:number double bonds (position). Throughout the specificationlinoleic acid is referred to as either linoleic acid or “LA”.

As used herein the term “arachidonic acid” refers to 5, 8, 11, 14eicosatetraenoic acid, which has a short hand designation of 20:4(n-6)and a molecular weight of 304.5. Throughout the specificationarachidonic acid is referred to as either arachidonic acid or “AA”. Itshould be noted that arachidonic acid can yield pro-inflammatoryprostaglandins. It also should be noted that arachidonic acid can beinvolved in enzymatic processes that result in beneficialanti-inflammatory lipid mediators such as lipoxins and anendocannabinoid that is anandamide (arachidonoylethanolamine).

As used herein the term “alpha-linolenic acid” refers to 9, 12, 15octadecatrienoic acid, which has a short hand designation of 18:3(n-3)and a molecular weight of 278.4. Throughout the specificationalpha-linolenic acid is referred to as either alpha-linolenic acid or“ALA”.

As used herein the term “gamma-linolenic acid” refers to 9, 6,12,-octadecatrienoic acid, which has a short hand designation of18:3(n-6) and a molecular weight of 278.4. Throughout the specificationgamma-linolenic acid is referred to as either gamma-linolenic acid or“GLA”.

As used herein the term “dihomogamma-linolenic acid” refers to 8, 11, 14eicosatrienoic acid, which has a short hand designation of 20:3(n-6) anda molecular weight of 306.5. Throughout the specification eicosatrienoicacid is referred to as either eicosatrienoic acid or “DGLA”.

As used herein the term “eicosapentaenoic acid” refers to5,8,11,14,17-eicosapentaenoic acid, which has a short hand designationof 20:5(n-3) and a molecular weight of 302.5. Throughout thespecification eicosapentaenoic acid is referred to as eithereicosapentaenoic acid or “EPA”.

As used herein the term “docosahexaenoic acid” refers to4,7,10,13,16,19-docosahexaenoic acid, which has a short hand designationof 22:6(n-3) and a molecular weight of 328.6. Throughout thespecification docosahexaenoic acid is referred to as eitherdocosahexaenoic acid or “DHA”.

As used herein, the term “ester” refers to any chemical compound derivedby reaction of an oxoacid (an organic acid that contains oxygen) with ahydroxyl compound, such as an alcohol. Esters are usually derived froman organic acid in which at least one hydroxyl (—OH) group is replacedby an —O-alkyl (alkoxy) group. Most commonly, esters are formed bycondensing a carboxylic acid with an alcohol. In one or moreembodiments, the esters of the present invention can be naturallyoccurring, or can be formed by reaction of a fatty acid with an alcohol.

As used herein, the term “amidoester” refers to any chemical compoundderived by reaction of an oxoacid with an amine. One or more embodimentsprovide that the reaction of a fatty acid with an amine provides anamidoester. Reference to the “ester form” can include the amidoester inaddition to the traditional ester. Reference to “reaction product” meansa resulting ester or amidoester that is formed by reaction of an acidwith an alcohol or amine, regardless of whether the ester or amidoesteris naturally-occurring or synthesized. If synthesized, the ester oramidoester can be prepared by various esterification methods known toone skilled in the art.

As used herein, the term “wax ester” refers to an ester of a fatty acidand a long-chain alcohol. Wax esters include, without limitation,beeswax and carnauba wax. Beeswax consists of C₄₀ to C₄₆ molecularspecies. Carnauba wax constitutes from C₁₆ to C₂₀ fatty acids esterifiedwith C₃₀ to C₃₄ long-chain alcohols to provide a C₄₆ to C₅₄ molecularspecies.

As used herein, the term “alcohol” refers to any organic compoundcontaining at least one hydroxyl functional group (—OH) bound to acarbon atom, that is usually bound to other carbon and hydrogen atoms;this includes, but is not limited to, acyclic alcohols; cyclic alcohols;primary, secondary, and tertiary alcohols; monohydric alcohols. Thealcohols include monohydric alcohols, which are alcohols containing asingle hydroxyl functional group. Monohydric alcohols include anycompound of the formula CH₃—(CH₂)_(z)—OH wherein z is from 0 to 5, andin some embodiments 0-2. Monohydric alcohols can especially includeethanol, CH₃—CH₂—OH.

As used herein, the term “resolvin” is an agent that is generated fromthe interaction between an omega-three polyunsaturated fatty acid suchas eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA),cyclooxygenase-II (COX-2) and an analgesic, such as aspirin. Resolvinsof the E series are derived from EPA, whereas resolvins of the D seriesare derived from DHA. Exemplary resolvins include resolvin E1 (RvE1),resolvin E2 (RvE2), resolvin D1 (RvD1), resolvin D2 (RvD2), resolvin D3(RvD3), resolvin D4 (RvD4), and combinations thereof.

As used herein, the term “protectin” or “neuroprotectin” is an agent,more particularly, a docosanoid (which is a signaling molecule made byoxygenation of 22-carbon essential fatty acids, especially DHA), that isderived from the polyunsaturated fatty acid docosahexaenoic acid (DHA).A “protectin” or “neuroprotectin” exerts potent anti-inflammatory andanti-apoptotic bioactivity at nanomolar concentrations in a variety ofexperimental models of brain and retinal diseases. An exemplaryprotectin includes protectin D1 (PD1).

As used herein, the term “lipoxin” refers to a series ofanti-inflammatory lipid mediators that are synthesized by the5-lipoxygenase pathway. Lipoxins are short-lived, endogenously produced,non-classic tetraene-containing eicosanoids, whose appearance ininflammation signals the resolution of inflammation. Lipoxins are alsoderived enzymatically from arachidonic acid, an omega-six fatty acid.Exemplary lipoxins include lipoxin A4 (LXA), lipoxin B4 (LXB4), andcombinations thereof.

As used herein, the term “prostaglandin” refers to one of a number ofhormone-like substances that participate in a wide range of bodyfunctions such as the contraction and relaxation of smooth muscle, thedilation and constriction of blood vessels, control of blood pressure,and modulation of inflammation. Prostaglandins are derived fromomega-three and omega-six fatty acids acid. There are three main typesof prostaglandins: Prostaglandin E1 (PGE1) and prostaglandin E3 (PGE3),which have anti-inflammatory properties, and prostaglandin E2 (PGE2),which promotes inflammation. PGE1, derived from dihomo-gamma-linolenicacid, is a potent vasodilator agent that increases peripheral bloodflow, inhibits platelet aggregation, and has many other biologicaleffects such as bronchodilation, and mediation of inflammation. PGE1 isimportant for lacrimal and salivary gland secretion and for T cellfunction. PGE2, derived from arachidonic acid, is released by bloodvessel walls in response to infection or inflammation and acts on thebrain to induce fever; PGE2 has also been used extensively as anoxytocic agent. PGE3, is formed via the cyclooxygenase (COX) metabolismof eicosapentaenoic acid. It is known that PGE3 lowers intraocularpressure.

As used herein, the term “retinoic acid” refers to a metabolite ofVitamin A (retinol) that mediates the functions of Vitamin A requiredfor growth and development. Retinoic acids have been shown to havestrong anti-inflammatory properties, in addition to their function assebostaticums. (see Plewig, G., et al., Archives of DermatologicalResearch, Vol. 270, No. 1, 89-94). Retinoic acids can include, withoutlimitation, 13-cis-retinoic acid.

As used herein, the term “endocannabinoid” refers to a class of organiccompounds found produced within the body that activate cannabinoidreceptors. Endogenous cannabinoids (“endocannabinoids”), when present intissues at elevated concentrations, provide anti-inflammatory andanalgesic effects. Endocannabinoids serve as intercellular lipidmessengers, signaling molecules that are released from one cell andactivating the cannabinoid receptors present on other nearby cells; theyuse retrograde signaling. Endocannabinoids are lipophilic molecules thatare not very soluble in water. Endocannabinoids can include, withoutlimitation, anandamide (arachidonoylethanolamine) and2-arachidonoylglycerol.

As used herein, the term “phospholipid” refers to any of variousphosphorous-containing lipids that are composed mainly of fatty acids, aphosphate group, and a simple organic molecule such as choline.Preferably, the phospholipids contain residues of one or more fattyacids that are omega-3 fatty acids, along with, as desired, omega-6fatty acids. Phospholipids are amphipathic in nature; that is, the polarend of a phospholipid is soluble in water (hydrophilic) and aqueoussolutions, while, the fatty acid end is soluble in fats (hydrophobic).In an aqueous environment, phospholipids combine to form a two-layerstructure (lipid bilayer) with the hydrophobic ends in the middle andthe hydrophilic ends exposed to the aqueous environment. Such lipidbilayers are the structural basis of cell membranes.

As used herein, the term “metabolite” refers to a compound that is theproduct of metabolism. A metabolite is formed as part of the naturalbiochemical process of degrading and eliminating compounds.

As used herein, the term “metabolically stable analog” refers to acompound that is a structural derivative of a parent compound (sometimesdiffering from the parent compound by a single element), or is acompound with similar properties to the parent compound. The analog isnot easily degraded, and, thus, is metabolically stable.

As used herein the term “CD11b+ infiltration” includes the increase inCD11b⁺ cells present in the center and periphery of the cornea followingdry eye induction.

As used herein the term “IL-1α or TNF-α expression” includes measuringRNA transcripts of IL-1α and TNF-α by quantitative real-time PolymeraseChain Reaction.

As used herein the term “inflammatory cytokines” includes, withoutlimitation, IL-1α and TNF-α.

Turning to the details of the disclosure, provided are processes ofmaking and using ocular products containing esterified anti-inflammatorylipid mediators, wherein the majority of the anti-inflammatory lipidmediator is present in an ester form. One or more embodiments providethat the compositions are substantially free of fatty acids. That is, insuch embodiments, the ocular products contain 10% by weight or less (or8%, or 6%, or 5%, or 4%, or 3%, or 2%, or even 1%) of the acid form ofthe anti-inflammatory lipid mediator. In a further embodiment, theocular products contain 1% by weight or less (or 0.8%, or 0.6%, or 0.5%,or 0.4%, or 0.3%, or 0.2%, or even 0.1%, or 0.05%, or 0.025%, or 0.01%)of the acid form of the anti-inflammatory lipid mediator. The esterifiedanti-inflammatory lipid mediators are esters of an acidanti-inflammatory lipid mediator. The esters may be formed by reactingthe anti-inflammatory lipid mediator with at least one monohydricalcohol or amine. Other embodiments allow for the ester to be formedfrom an amine. Desirable anti-inflammatory lipid mediators includeomega-three and/or omega-six fatty acids, resolvins or a metabolicallystable analog, protectins or a metabolically stable analog, lipoxins ora metabolically stable analog, prostaglandins or a metabolically stableanalog, retinoic acids, endocannabinoids, and phospholipids.Inflammation is a component of dry eye. There is a need to deliveractive candidates, known to mitigate inflammation, in forms that are notassociated with initial discomfort (acute ocular discomfort) uponadministration to the eye, while providing long-term benefits to theeye.

One or more embodiments provide that the ester is provided in atherapeutically effective amount. That is, the ester is present in anamount sufficient to provide a beneficial effect to the ocular area,including but not limited to the ocular surface, the back of the eye,tear formation and stability. A therapeutically effective amount ofester can deliver an appropriate amount of anti-inflammatory lipidmediator that imparts a benefit to the ocular environment.

In the free fatty acid formulations (for example, alpha-linolenic acidemulsions) of the prior art (e.g., those compositions disclosed in U.S.Patent Application Pub. 20070265341 (Dana et al.)), discomfort uponinstillation to the eye has been found. A change in the concentration ofsurfactants (mostly Tween-80, from 2.5% to 0.25%) or the use ofadditional surfactant(s) (such as the amphoteric monateric surfactant)did not result in improved comfort upon instillation. The currentinvention seeks to avoid or remedy the discomfort issue by making theessential fatty acid non-ionic, i.e., using the esterified counterpartof the molecule.

Anti-inflammatory lipid mediators, such as polyunsaturated fatty acids,resolvins or a metabolically stable analog, protectins or ametabolically stable analog, lipoxins or a metabolically stable analog,prostaglandins or a metabolically stable analog, retinoic acids,endocannabinoids, and phospholipids are desirable ingredients of ocularproducts for use in treating such ocular conditions as inflammation, dryeye and/or dryness symptoms, and meibomian gland dysfunction. It hasbeen discovered that the use of esterified anti-inflammatory lipidmediators, when the majority of the anti-inflammatory lipid mediator ispresent in the ester form, results in an ocular product that greatlyimproves initial comfort upon contact with or administration to theocular surface. Generally, the esterified anti-inflammatory lipidmediator is a reaction product of an acid anti-inflammatory lipidmediator and an alcohol or an amine.

Such esterified anti-inflammatory lipid mediators may also be useful ina rewetting drop, in some instances unpreserved, or may be associatedwith a contact lens, such as a silicone hydrogel, whereby the lens maybe treated with a mixture of the esterified anti-inflammatory lipidmediators. The esterified anti-inflammatory lipid mediators can beincorporated into the contact lens using various methods, for example,incorporation can occur during the lens extraction or hydration processor a combination thereof.

Such a characteristic is not offered by previous uses of fatty acidsand/or fatty acid oils. The esterified anti-inflammatory lipid mediatorscan be combined with an aqueous delivery system—for desired ophthalmiccompositions.

Esterified anti-inflammatory lipid mediators, when the majority of theanti-inflammatory lipid mediator is present in the ester form, have theadvantage of targeting the inflammatory component of the dry eye disease(which perpetuates dry eye disease) and are less likely to cause initialdiscomfort at a wider concentration range. Upon contact with and uptaketo the cells of the ocular surface, and without intending to be bound bytheory, it is thought that esterified anti-inflammatory lipid mediators,such as esterified polyunsaturated fatty acids, esterified resolvins ora metabolically stable analog, esterified protectins or a metabolicallystable analog, esterified lipoxins or a metabolically stable analog,esterified prostaglandins or a metabolically stable analog, esterifiedretinoic acids, endocannabinoids, and phospholipids, undergo hydrolysisand return to their acidic anti-inflammatory lipid mediator state alongwith the alcohol that was used in forming the ester.

Turning to the esters of polyunsaturated fatty acids such as omega-threeand omega-six fatty acids, the reaction of carboxylic acids and alcoholsor acetates will produce esters. In general terms, the following fattyacid derivatives such as esters (Ia) and other functionalities such asamides (Ib) are desirable for their stability and improved initial eyecomfort:CH₃—CH₂—CH═CH—(CH₂—CH═CH)_(n)—(CH₂)_(x)—CO—O—R  (Ia)CH₃—CH₂—CH═CH—(CH₂—CH═CH)_(n)—(CH₂)_(x)—CO—NH—R  (Ib)

Such derivatives are then expected to be converted back to theiroriginal fatty acid structure (II):CH₃—CH₂—CH═CH—(CH₂—CH═CH)_(n)—(CH₂)_(x)-acid{—CO—OH}  (II)

Once in the ocular environment and/or incorporated into the lipid layeror cell membrane lipid bilayer to carry-on their tear film stabilizationeffect and/or anti-inflammatory effect.

The ranges of n, x and R can fall within the following ranges: n: 2-5;x: 2-7; R: ophthalmologically compatible leaving group included, but notlimited to:

—(CH₂)_(y)CH₃, where y is 0, 1 or above. In some embodiments y isbetween 0 and 5, or even 0 and 3, with y=1 being preferred.

Specifically, without limitation, the esterified anti-inflammatory lipidmediator comprises an esterified omega-three fatty acid, wherein theomega-three fatty acid is selected from the group consisting of:alpha-linolenic acid, stearidonic acid, eicosatetraenoic acid,eicosapentaenoic acid, docohexaenoic acid, docosapentaenoic acid (DPA),tetracosapentaenoic acid, and tetracosahexaenoic acid (Nisinic acid),derivatives, metabolites, and mixtures thereof. Upon esterification, themajority of the esterified anti-inflammatory lipid mediator is presentin the ester form of the omega-three fatty acid.

Specially, esterified omega-three fatty acids can be selected from thefollowing non-limiting examples: ethyl linolenate (alpha-linolenic acidethyl ester (ALA-EE); stearidonic acid ethyl ester and stearidonic acidpropyl ester; eicosatetraenoic acid ethyl ester and eicosatetraenoicacid propyl ester; eicosapentaenoic acid ethyl ester andeicosapentaenoic acid propyl ester; and docohexaenoic acid ethyl esterand docohexaenoic acid propyl ester.

The anti-inflammatory lipid mediator is reacted with a monohydricalcohol or an amine to form the desired ester form of theanti-inflammatory lipid mediator. Monohydric alcohols containing asingle hydroxyl functional group. Suitable monohydric alcohols includeup to 5 carbon atoms. In one embodiment, the monohydric alcohols havethe formula CH₃—(CH₂)_(z)—OH wherein z is 0 to 5. In another embodimentthe monohydric alcohols are selected from methanol and ethanol. Inanother embodiment the monohydric alcohol is ethanol.

Suitable amines include primary and secondary amines having up to 6carbon atoms. The amines may be linear, branched or cyclic. In oneembodiment suitable amines include amines having the formulaCH₃—(CH₂)_(z)—NH₂ wherein z is 0 to 5, and preferably 0 or 1.

The inflammatory lipid mediator and monohydric alcohol are reacted underester forming conditions. Suitable catalysts are known in the art andinclude acids, bases, carbodiimide, and the like. The esterification andamidation reactions can take place at room temperature (typically in therange of about 19-25° C.) without much need to go higher and ambientpressure, temperatures can be brought to higher ranges (about 25° C. to80° C.) in order to accelerate the time to reaction completion.

It should be noted that an esterified anti-inflammatory lipid mediatordoes not take the form of naturally occurring oils including sunfloweroil, sesame oil, castor oil, linseed oil, and the like. It should befurther noted that the esterified anti-inflammatory lipid mediators ofthe present invention are not wax esters as they are formed fromalcohols having short carbon chains (six or less and in some embodiments3 or less carbon atoms).

Mixtures may include omega-three and omega-six fatty acid esters atdesired ratios. In one or more embodiments it is desirable to provide acomposition which will, upon hydrolysis of the ester, provide a balanceof omega-three fatty acid:omega-six fatty acid in the eye to about 1:1.In other embodiments it is desirable to provide ophthalmic compositionswhich have ratios of the omega-three fatty acid:omega-six fatty acidupon hydrolysis, in the range of about 10:about 1 to no less than about1:about 1 and from about 5:1 to about 1:1, from about 4:1 to about 1:1,from about 3:1 to about 1:1, from about 2:1 to about 1:1, about 1:1,about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about8:1, about 9:1, or about 10:1. The ratio is based on the total amount ofeach class of omega fatty acids.

Mixing of the ophthalmic compositions can be done under asepticconditions, or under ambient conditions and then sterilized. Temperaturecan range widely and the reactions may be performed under ambientconditions of temperature and pressure.

In addition to the usefulness of esterified anti-inflammatory lipidmediators as ingredients in ophthalmic compositions, includingre-wetting drops, multipurpose solutions, cleaning and storing solutionsand in contact lenses themselves, such materials are also candidates fortheir inclusion in lens packing solution. Lenses may be packaged withesterified anti-inflammatory lipid mediators in formulations and/oremulsions or may be hydrated in such materials as dissolved inappropriate solvent(s), followed by equilibration of the lens in packingsolution.

Other ophthalmic compositions include lens care solutions such asmultipurpose solutions, preparations, gels, ointments, emulsions, andophthalmic products such as strips, inserts or punctal plugs or anyproduct coming into contact with the ocular surface.

In one embodiment, the esterified anti-inflammatory lipid mediators areprovided in an aqueous delivery system. Aqueous delivery systems arewater based systems, which can be instilled directly in the eye, or maybe used to condition, store, or clean ophthalmic devices which areplaced in the ocular environment. Examples of aqueous delivery systemscan include one or more of the following: packing solutions, storingsolutions, cleaning and care solutions, multipurpose solutions,conditioning solution and ophthalmic drops. The aqueous delivery systemsmay also include known components, such as one or more of emulsifiers,chelant agents, or stabilizers, surfactants, wetting agents,antioxidants, tonicity adjusting agents, preservatives, osmoprotectionagents, combinations thereof, and the like.

The packaging solution may be any water-based solution including thatwhich is used for the storage of contact lenses. The esterifiedanti-inflammatory lipid mediators are dispersed in the packagingsolution. Typical solutions include, without limitation, salinesolutions, other buffered solutions, and deionized water. The preferredaqueous solution is saline solution containing salts including, withoutlimitation, sodium chloride, sodium borate, sodium phosphate, sodiumhydrogenphosphate, sodium dihydrogenphosphate, or the correspondingpotassium salts of the same. These ingredients are generally combined toform buffered solutions that include an acid and its conjugate base, sothat addition of acids and bases cause only a relatively small change inpH. The buffered solutions may additionally include2-(N-morpholino)ethanesulfonic acid (YMS), sodium hydroxide,2,2-bis(hydroxymethyl)-2,2′,2″-nitrilotriethanol,n-tris(hydroxymethyl)methyl-2-aminoethanesulfonic acid, citric acid,sodium citrate, sodium carbonate, sodium bicarbonate, acetic acid,sodium acetate, and the like and combinations thereof. Preferably, thesolution is a borate buffered or phosphate buffered saline solution.

To form the packaging solution, at least one surfactant or emulsifieralong with any additional ingredients are combined with the water-basedsolution, stirred, and dissolved or dispersed. The pH of the solutionpreferably is adjusted to about 6.2 to about 7.5. The lens to be storedin the packaging solution of the invention is immersed in the solutionand the solution and lens placed in the package in which the lens is tobe stored. Alternatively, the solution may be placed into the packageand the lens then placed into the solution. Typically, the package isthen sealed by any convenient method, such as by heat sealing, andundergoes a suitable sterilization procedure.

The surfactants suitable for use in the invention are of any suitablemolecular weight, preferably about 200 to about 1,000,000, morepreferably about 1000 to about 18,000. Useful surfactants have ahydrophile-lipophile balance (“HLB”) of about 10 to about 30, preferablyabout 15 to about 25, more preferably about 15 to about 23.

Any of the known surfactants fitting the aforementioned criteria may beused provided that the surfactant is compatible, in terms of solubility,in the solution with which it is used. Thus, suitable surfactantsinclude, without limitation, cationic, ionic, non-ionic surfactants, andcombinations thereof. However, the use of a lens packaging solutioncontaining cationic and ionic surfactants may cause eye irritation.Therefore, preferably the surfactant is a non-ionic surfactant.

Suitable non-ionic surfactants include, without limitation, polyethyleneglycol esters of fatty acids, such as polysorbate 20, 60 or 80, allavailable as TWEEN® surfactants, alkanolamides, amine oxides,ethoxylated alcohols and acids, and surfactants having one or morepoly(oxyalkylene) chains, such as poloxamine surfactants (asurface-active agent that removes lipid and environmental debris fromthe lenses; polyalkoxylated block polymers of ethylene diamine) orpoloxamer surfactants (any of a series of nonionic surfactants of thepolyoxypropylene-polyoxyethylene copolymer type, used as surfactants,emulsifiers, stabilizers, and food additives), and the like, andcombinations thereof. Preferably, the surfactant is a polysorbate orpoloxamer surfactant. Poloxamer surfactants are commercially availableunder the name PLURONIC200 that are polyoxyethylene-polyoxypropylenenon-ionic surfactants having polyoxyethyl hydrophilic group ends thatmake up about 10 to about 80 percent by weight of the molecule. Althoughany of the PLURONIC® surfactants are preferred, particularly preferredfor use in the invention is PLURONIC® 127, which is about 70 percent byweight ethylene oxide and has a molecular weight of about 12,000 toabout 15,0000.

The surfactant may be combined with any known active and carriercomponents useful for lens packaging solution or for a rewetting drop.Suitable active ingredients for lens packaging solutions include,without limitation, antibacterial agents, anti-dryness agents, such aspolyvinyl alcohol, polyvinylpyrrolidone, and dextran, tonicity agents,and the like, and combinations thereof.

Suitable wetting agents, along with viscosity enhancers include, withoutlimitation: methyl gluceth-20 (sold under the trade name, for example,Glucam E20), carboxymethylcellulose, dextran 70, gelatin,hydroxymethylcellulose, hydroxypropyl methylcellulose,hydroxypropylethylcellulose, hydroxypropyl cellulose, methylcellulose,PEG, propylene glycol, polyvinyl alcohol (PVA), polyvinylpyrrolidone(PVP), Carbomer, polymethylvinylethermaleic anhydride, hyaluronic acid,xanthan gum, and polyacrylicacid.

Suitable antioxidants used in this invention include, withoutlimitation, hindered phenols, catechols, tocopherols, carotenoids,hyaluronic acid, lutein, or any species that can scavenge free radicals.Antioxidants are molecular species that inhibit oxidative damage ofother chemicals through redox chemical reactions. These reactionstypically transfer electrons for a molecule species to an oxidantmolecule. These can include free radicals, which can cause chainreactions. In simplest terms, antioxidants are reducing agents. Examplesof antioxidants include, without limitation: Vitamin E, Vitamin C, betacarotene (which is converted to Vitamin A), and peroxidases, and otheragents which can inhibit the formation of free radicals, e.g., chelants,EDTA, diethylene triamine pentaacetic acid (DTPA), N,N-bis[carboxymethyl]glycine (NTA), and the like.

In some embodiments, Vitamin E is added to a solution comprising theesterified anti inflammatory lipid mediator.

In another embodiment the composition of the present invention isincorporated into an ophthalmic device such as a contact lens or, moreparticularly, a silicone hydrogel contact lens. In this embodiment theesterified anti-inflammatory lipid mediators, wherein the majority ofthe anti-inflammatory lipid mediator is present in the ester form, maybe incorporated into the lens in a number of ways, including but notlimited to incorporating into the reaction mixture from which the lensis polymerized, contacting the lens with a solution comprising theesterified anti-inflammatory lipid mediators either before during orafter packaging. For example the esterified anti-inflammatory lipidmediators may be included in the extraction, hydration or storagesolution during the manufacture of the lens or may be included in asolution which is contacted with the contact lens by the lens wearer. Inone embodiment the solution swells the lens, which allows for enhanceduptake of the esterified anti-inflammatory lipid mediators. Inembodiments where the esterified anti-inflammatory lipid mediator isincorporated into the reaction mixture, the esterified anti-inflammatorylipid mediator may be added to the reaction mixture as a separatecomponent, or may be pre-reacted with the alcohol group on at least oneof the reactive components.

In some embodiments, the present invention comprises ophthalmiccompositions comprising at least one esterified omega-three fatty acid.In some embodiments, the present invention comprises ophthalmiccompositions comprising at least one esterified omega-six fatty acid. Insome embodiments, the present invention comprises ophthalmiccompositions comprising at least one esterified omega-six fatty acid andat least one esterified omega-three fatty acid.

It is a benefit of the present invention that the esterifiedanti-inflammatory lipid mediators are hydrolytically stable at neutralpH, and do not hydrolyze during storage in the pH neutral ophthalmiccomposition and sterile preparations of the present invention. Thismeans that the ophthalmic solutions and sterile preparations do notcause stinging when instilled in the eye. Upon contact with the cellularmembranes and/or transport into the cells of the ocular surface, andwithout intending to be bound by theory, it is thought that esterifiedanti-inflammatory lipid mediators, such as esterified polyunsaturatedfatty acids, esterified resolvins or a metabolically stable analog,esterified protectins or a metabolically stable analog, esterifiedlipoxins or a metabolically stable analog, esterified prostaglandins ora metabolically stable analog, esterified retinoic acids,endocannabinoids, and phospholipids, undergo hydrolysis and return totheir acidic anti-inflammatory lipid mediator state along with thealcohol that was used in forming the ester.

The amounts of the esterified anti-inflammatory lipid mediator can bestated as a percentage of the total composition or as a percentage ofthe solution used in a processing step such as a lens hydration step(part of the lens making process that can result in the incorporation ofthe material into the device). The percentage of esterifiedanti-inflammatory lipid mediator can be determined by any method, butcan, for example, be determined by dividing the weight of theanti-inflammatory lipid mediator by the total weight of the ophthalmiccomposition or device. The percentage of any component of the ophthalmiccomposition can be determined in a similar manner.

The amount of esterified anti-inflammatory lipid mediator which may bepresent in the ophthalmic compositions or devices of the presentinvention include from about 0.025 weight % to 5.0 weight % based uponall the components in the ophthalmic composition. When the ophthalmiccomposition is a rewetting drop, the esterified anti-inflammatory lipidmediator is present in an amount from about 0.025 weight % to 0.5 weight% based upon all of the components in the composition, and the acidcontent can be no more than 0.1 weight % (or 0.075, or 0.05, or 0.025,or even 0.01 weight %). When the ophthalmic composition is incorporatedonto a contact lens, the esterified anti-inflammatory lipid mediator ispresent in an amount from 0.025 weight % to 5.0 weight % based upon allof the components in the composition, and the acid content can be nomore than 1 weight % (or 0.75, or 0.5, or 0.25, or even 0.1 weight %).

In some embodiments, the invention is directed to the topicalapplication of a composition comprising an esterified anti-inflammatorylipid mediator (e.g., esterified ALA) as an effective therapeuticstrategy to decrease ocular surface inflammation. As discussed hereinthe inflammation of the ocular surface can be seen in, for example, dryeye syndrome and other inflammatory ocular conditions including, but notlimited to, both anterior segment/front of the eye conditions and backof the eye conditions (e.g., meibomian gland dysfunction, blepharitis,atopic keratoconjunctivitis, contact lens related dry eye, Sjögren'ssyndrome, uveitis, macular degeneration, and a wide range of otherconditions).

In another embodiment, the invention is directed to the topicalapplication of a composition comprising an esterified anti-inflammatorylipid mediator (e.g., esterified ALA) as an effective strategy toimprove tear film function or tear film stability. Without intending tobe bound by theory, it is thought that the esterified anti-inflammatorylipid mediator improves the interaction between the lens and the tearfilm and/or the lids.

The present invention can also be administered to an individual that hasbeen identified in need thereof of a composition described herein. Theindividual can be in need thereof, if the individual has been identifiedas suffering or having the condition of dry eye syndrome or one of theother inflammatory ocular conditions identified above. One in skill inthe art would know how to identify the individual in need of a treatmentfor dry eye syndrome.

The present invention can also be administered to an individual tomitigate at least one sign and/or symptom of dry eye, or to provideosmoprotection to an individual in need thereof.

Without intending to be bound by theory, it is thought that when theanti-inflammatory lipid mediator composition is loaded onto a contactlens for delivery to the eye during contact lens wear, by virtue of itsanti-inflammatory properties and the benefit provided to the tear film,the anti-inflammatory lipid mediator can be held on to the eye via thecontact lens long enough to be delivered efficiently to the eye in orderprovide relief to individuals suffering from dry eye or otherinflammatory ocular conditions.

Epithelial Construct Method

A model was developed to allow for the prediction of acute oculardiscomfort obtained with fatty acid rewetting drops. The model uses acorneal epithelium tissue construct (e.g. HCE construct manufactured bySkinEthic). This tissue construct is formed by a full thickness cornealepithelium tissue reconstituted in vitro whereby human cornealepithelial cells were cultured at the air liquid interface in achemically-defined medium, resulting in the formation of a cornealepithelial tissue devoid of stratum corneum and resemblinghistologically the mucosa of the human eye (Nguyen, D. H., et al.,Three-dimensional construct of the human corneal epithelium for In Vitrotoxicology. In: Alternatives Toxicological Methods. Boca Raton, Fla.,CRC Press ed Katz S A and Salem H, 2003. Chapter 14: p. 147-159). Thetissue is approximately 70 microns thick and has been shown to expressthe relevant markers of the fully differentiated corneal epithelialtissue (Nguyen et al, 2003).

The construct was further characterized, whereby dose-dependent effectsof the common ophthalmic preservative benzalkonium chloride (BAK) wereevaluated in terms of both cell viability impact and inflammatorycytokine release/cell activation. It was shown that the cell viabilityresponse in the model was consistent with the known in vivo effects ofthe BAK within the range of concentration that was evaluated. Thecytokine endpoint (quantification of II-1 alpha release in the tissueculture media) allowed sensitive evaluations of the effects of theophthalmic preservative on cell activation considering it allowed todiscriminate non-cytotoxic concentrations of BAK (0.001 and 0.01%).

The esterified anti-inflammatory lipid mediator dose-responses were thenevaluated in the construct, whereby 30 microliters of the rewetting dropwas topically applied to the tissue for 8 or 24 hours, and cellviability was then quantified along with a subset of selected potentialbiomarkers within the tissue culture media. In vitro endpoints wereprobed for their correlation with the in vivo response (comfort uponinstillation) in order to identify AOD predictors and define the (acuteocular) comfort prediction model.

It was determined that the 8 hour incubation period was the mostrelevant for optimal correlation between the in vitro and the in vivoresponses. Using the 8 hour exposure condition, a very good correlationwas obtained between the in vivo response of comfort upon instillation,measured immediately upon instillation, using a scale of 0 to 50, with50 being the most comfortable score with a comfort rating of“Excellent,” with a score of approximately 42 having a comfort rating of“Good,” with a score of approximately 33 having a comfort rating of“better than satisfactory” (i.e., above average), with a score ofapproximately 26 having a comfort rating of “satisfactory” (i.e.,average), with a score of approximately 18 having a comfort rating of“less than satisfactory” (i.e., below average), with a score ofapproximately 8 having a comfort rating of “poor,” and 0 being the leastcomfortable score with a comfort rating of “very poor,” and thefollowing in vitro endpoints.

Table 1 shows the cell viability as assessed using the MTT assay.

TABLE 1 Cell Viability (% of PBS Comfort Score Average control) 45 +/−7  105 +/− 2  41 +/− 9  99 +/− 6 27 +/− 14 65 +/− 6 5 +/− 7  7 +/− 1

Table 2 shows II-1 alpha release in the tissue culture media.

TABLE 2 Il-1 Average (% of PBS Comfort Score Average response) 45 +/− 7 443 +/− 20 41 +/− 9  709 +/− 37 27 +/− 14 1276 +/− 109 5 +/− 7 3374 +/−283

Table 3 shows II-1 Ra release in the tissue culture media.

TABLE 3 Comfort Score Il-1 Ra Average (% of PBS Average response) 45 +/−7  123 +/− 3  41 +/− 9  235 +/− 6  27 +/− 14 386 +/− 25 5 +/− 7 917 +/−22

The desired outcome of in vitro response was defined based on the invitro-in vivo correlations. The undesirable comfort score correspondedto a score equal to or below 25, which translated into an undesirablecell viability in vitro endpoint of 60% or below, per in vivo-in vitrocorrelations. The most desirable comfort score was 35 or above, whichtranslated into a cell viability value of 80% or above, per in vivo-invitro correlations.

The model was further confirmed by showing that the alpha linolenic acidethyl ester, which had shown a beneficial in vitro response in themodel, also showed a beneficial response on the eye (i.e. lack of acuteocular discomfort).

The invention is now described with reference to the following examples.Before describing several exemplary embodiments of the invention, it isto be understood that the invention is not limited to the details ofconstruction or process steps set forth in the following description.The invention is capable of other embodiments and of being practiced orbeing carried out in various ways.

EXAMPLES Example 1

Topical compositions were formed by adding ethyl linolenate(alpha-linolenic acid ethyl ester (ALA-EE)), an esterifiedanti-inflammatory lipid mediator (AILM) having the formula:

to a packing solution/aqueous-based delivery system containing varioussurfactants/emulsifiers, wetting agent, and a chelant and antioxidant(s)followed by mixing at high shear rate, to provide an emulsion suitablefor topical application. Table 4 summarizes the ingredients ofcompositions.

TABLE 4 Example Example 1A 1B Example 1C Example 1D Modified packing99.3 94.8 94.8 94.9 solution^(a) (wt %) Glucam E20 (wt %) 0.25 2.5 2.52.5 Tween-80 (wt %) 0.25 2.5 2.5 2.5 ALA-EE (wt %) 0.20 0.20 0.20 0.10Vitamin E (wt %) 0.03 0.03 0.03 0.03 ^(a)= 0.4 wt % boric acid, 0.2 wt%, sodium borate, 0.5 wt % sodium chloride, 0.01 wt %Diethylenetriaminepentaacetic Acid (DTPA)

Example 2

Comparative

Topical compositions were formed by adding alpha-linolenic acid (ALA) toa packing solution/aqueous-based delivery system containing varioussurfactants/emulsifiers, wetting agent, and a chelant and antioxidant(s)followed by mixing at high shear rate, to provide an emulsion suitablefor topical application. Table 5 summarizes the ingredients ofcompositions.

TABLE 5 Example Example 2A 2B Example 2C Example 2D Modified packing99.3 99.4 94.8 94.9 solution^(a) (wt %) Glucam E20 (wt %) 0.25 0.25 2.52.5 Tween-80 (wt %) 0.25 0.25 2.5 2.5 ALA (wt %) 0.20 0.10 0.20 0.10Vitamin E (wt %) 0.03 0.03 0.03 0.03 ^(a)= 0.4 wt % boric acid, 0.2 wt%, sodium borate, 0.5 wt % sodium chloride, 0.01 wt %Diethylenetriaminepentaacetic Acid (DTPA)

Example 3

Testing

The compositions of Examples 1A and 1B and Comparative Example 2A weretested in vitro as follows. A transepithelial permeability in vitromodel (TEP test) was used to assess fluorescein leakage (or increasedtransepithelial permeability) with the free acid in comparison with theester form of the alpha linolenic acid. Results are provided in Table 6,which shows that the ALA ethyl ester at high concentration (0.2%)behaved similarly to the vehicle (packing solution with Tween andGlucam) in terms of the effect on transepithelial permeability, whereasthe ALA fatty acid showed a significant increase in fluorescein leakageat the same concentration, relative to the controls, that was consistentwith the clinical observation of discomfort upon instillation. Thissuggests that the ester version of the omega-three fatty acid will havean improved tolerability profile upon instillation compared to the useof the free acid when using concentrations as high as 0.2%.

TABLE 6 Fluorescein leakage (Fluorescence units) HBSS Buffer 936 +/− 280Packing Sol'n 847 +/− 373 Vehicle 0.25% TW^(c) and Glcm^(d) 572 +/− 514Example 2A 14004 +/− 5544  Example 1A 545 +/− 462 Example 1B 720 +/− 531^(c)TW is Tween-80 ^(d)Glcm is Glucam E20

Example 4A

Testing

The compositions of Examples 1C, 1D and Comparative Examples 2C, 2D weretested in vitro as follows. A corneal epithelial tissue model consistingof a multi-endpoint assay system whereby the cell viability wasevaluated in addition to the measurement of biomarkers or specificcytokines which were previously shown (see tables 1-3 with correspondingcorrelation curves) to correlate with the in vivo data of subjectivediscomfort (upon instillation). Results for 8 hours are provided inTable 7A, which shows that the ethyllinolenate (ALA-EE) had 10 timesless impact on the in vitro endpoint (after 8 hour exposure) than thelinolenic acid (ALA), suggesting that ALA-EE may be used atconcentrations of up to 0.2% or higher without inducing initialdiscomfort upon instillation.

TABLE 7A Cell viability Form Acid/Ester (% of PBS ctrl) of AILMConcentration Vehicle 105 +/− 2  N/A N/A Example 1C 70 +/− 8 Ester 0.2wt % Example 1D 81 +/− 1 Ester 0.1 wt % Example 2C Comparison  7 +/− 1Acid 0.2 wt % Example 2D Comparison 65 +/− 6 Acid 0.1 wt %

Example 4B

Testing

The compositions of Example 1C (ALA-EE 0.2%) and Comparative Example 2C(ALA 0.2%) were tested in vitro using SkinEthic 0.1 cm² human cornealepithelium tissue constructs (HTS constructs) and 24 hours exposure timepoint. The results are shown in Table 7B, below.

Overall, the in vitro data of Examples 3, 4A, and 4B indicate that theester version of the omega three fatty acid is much less likely to causeinitial discomfort upon instillation, compared to the free acid and thatits use would offer the advantage to allow the use of a higherconcentration (for efficacy) without any risk of causing initialdiscomfort response.

TABLE 7B Acid/Ester Cell viability II-1 alpha Form of Concen- (% of PBSctrl) (pg/ml) AILM tration Vehicle 75.75 +/− 7.17 152.75 +/− 20.15 N/AN/A Example  3.73 +/− 1.38  809.25 +/− 240.77 Acid 0.2 wt % 2C Com-parative Example 87.00 +/− 7.63 112.48 +/− 0.04  Ester 0.2 wt % 1C PBS100.00 +/− 5.98  31.63 +/− 4.00 N/A N/A

Example 5

Preliminary non-dispensing clinical studies were conducted to furtherassess the use of omega-three fatty acid compositions. A 0.05 wt. %concentration ester version of an omega-three fatty acid was instilledon three “contact lens induced dry eye” subjects (each with at least onesign and one symptom of contact lens induced dry eye) without any acuteocular discomfort or physiological findings. Similarly a 0.2 wt. %concentration ester version of the omega-three fatty acid was instilledon four “contact lens induced dry eye” subjects without any acute oculardiscomfort or physiological findings.

Example 6

Omega-three ester material (notably, the alpha linolenic acid ethylester (ALA-EE) or ethyl linolenate) was loaded into a contact lens forits slow delivery on the eye during contact lens wear. By virtue of itsanti-inflammatory properties and the benefit it might provide to thetear film, the omega-three fatty acid (ester) can be held on the eye viathe contact lens long enough to be delivered efficiently to the eye inorder to provide relief in subjects suffering from dry eye or otherinflammatory conditions. Conditions targeted by this material includeboth anterior segment/front of the eye conditions and back of the eyeconditions, whereby the slow delivery of the omega-three fatty acid(ester) may provide protection against diseases such as maculardegeneration or may delay the onset of the condition, by virtue of thematerials' antioxidant and anti-inflammatory properties.

It is believed that the use of the ester form of the omega-three fattyacid derived from reaction with a monohydric alcohol provides thebenefit of improving bioavailability and allows for an on-eye release ofthe acid form upon encounter with the ocular tissue esterases. In thefollowing examples, the omega-three ester material was the linolenicacid ethyl ester or ethyl linolenate.

ALA-EE was dissolved in isopropanol (IPA) at the concentration of 0.05or 0.5%. Silicone hydrogel contact lenses (senofilcon, commerciallyavailable as ACUVUE® OASYS® with HYDRACLEAR® Plus lenses) were thenallowed to soak in the ALA-EE-containing IPA for up to 1 hour at roomtemperature, followed by soaking in ALA-EE containing IPA/packingsolution (70% IPA/30% packing solution) for up to 1 hour, rinsing inpacking solution, and packaging of lenses in the packing solution.ALA-EE uptake by the contact lenses was quantified by gaschromatography. No quantifiable free acid form was associated with thelenses, showing that the ALA-EE did not hydrolyze to the free acid formunder these conditions (Table 8).

ALA-EE uptake was 54 and 611 micrograms per lens using 0.05 and 0.5%ALA-EE solution in IPA, respectively (Table 6).

TABLE 8 % ALA-EE in ALA-EE IPA solution (μg/lens) ALA (μg/lens) Example6A 0.05 53.7 +/− 3.8 BDL* Example 6B 0.5 610.6 +/− 6.6  BDL* *BDL: belowdetection limit.

Example 7

Comparative

A comparative silicone hydrogel contact lens that incorporated alphalinolenic acid (ALA), rather than the ester, was prepared following thesame protocol as Example 6. The uptake of ALA by the contact lenses was23 and 300 micrograms per lens, when using a 0.05% and 0.5% ALA solutionin IPA, respectively (see Table 9).

TABLE 9 % ALA in IPA solution ALA (μg/lens) Example 7A 0.05 22.9 +/− 1.6Example 7B 0.5 299.0 +/− 16.5

Example 8

Testing

The evaluation of the ALA and ALA-EE lenses of Examples 6 and 7 in areconstituted human corneal epithelium construct showed a favorableinteraction with the ocular surface tissue following a 24 hour exposure,in terms of the absence of impact on cell viability. The cell viabilitywas not reduced relative to phosphate buffered saline (PBS) negativecontrol (Table 10).

TABLE 10 Cell viability (% of PBS ctrl) Example 6B 105 +/− 2 Example 7B 95 +/− 7 Control (no treatment) 110 +/− 1 PBS ctrl 24 hour 100 +/− 9

In Examples 4A and 4B, it was shown that the ester form of theomega-three fatty acid (ethyl linolenate) allowed the use of a higherconcentration of the material within a rewetting drop emulsion withoutcausing an initial discomfort.

In this example, it was demonstrated that the ester form of theomega-three fatty acid (ethyl linolenate) can be loaded at higherquantities than the free acid form using the same concentration of theraw material. Lenses incorporating such high amounts of the ester formof the omega-three fatty acid did not yield any loss in cell viability,suggesting acceptable ocular tolerability of the product.

Other methods of incorporation of the ethyl linolenate included the useof propylene glycol hydration (ALA-EE was solubilized into propyleneglycol instead of IPA for the hydration/soaking of the lenses) and itwas verified that no major changes in lens parameters were observed.

Example 9

The alpha linolenic acid ethyl ester (ALA-EE) was dissolved in propyleneglycol (PG) at concentrations of 0.05 and 0.5%, and silicon hydrogellenses (Narafilcon B lenses (ACUVUE® TrueEye®)) were hydrated or soakedin the ALA-EE-containing propylene glycol solution for one hour at roomtemperature, followed by rinsing in DI water and repackaging of lensesin the packing solution. The lenses were packaging in borate bufferpacking solution with 50 ppm methyl cellulose and autoclaved at 121° C.for 19 minutes. ALA-EE was quantified by gas chromatography.

Under these conditions and using the narafilcon B substrate, ALA-EEuptakes were 91 and 721 micrograms per lens using 0.05 and 0.5% ALA-EEsolution, respectively, in IPA, respectively (Table 11). There was nodetectable hydrolysis of the ester to the acid under these conditions.

TABLE 11 % ALA-EE in ALA-EE PG solution (μg/lens) ALA (μg/lens) Example9A 0.025 51.7 +/− 3.7 BDL* Example 9B 0.05  91.0 +/− 10.6 BDL* Example9C 0.5 721.5 +/− 39.3 BDL* *BDL: below detection limit.

There was no change in water content or lens diameter under theseconditions.

Example 10

Testing

The release of ALA-EE from selected lenses of Example 9 was assessedboth in vitro and in vivo (rabbit study), In vitro release in 1 ml of2.5% Tween-80 in packing solution showed that 50% of the ALA-EE wasreleased from a lens according to Example 9A within 3 days.

For in vivo testing, an animal study was conducted to evaluate oculartolerability of the product, and in particular, to assess in vivorelease rates. In vivo release in rabbit eyes showed a relatively slowrelease of the ALA-EE over time. With respect to a lens according toExample 9B, 43% of the ALA-EE was released within the first 12 hours,with an average of 3 micrograms per hour after the first hour. Over thecourse of one day, approximately 40 to 50 micrograms was released fromthe narafilcon B lens.

Examples 11-12

ALA-EE was dissolved in propylene glycol (PG) at concentrations of 0.05and 0.1%, and silicone hydrogel lenses (narafilcon B lenses (ACUVUE®TrueEye®)) were soaked at ambient temperature in the PG-ALA-EE solutions(200 lenses per liter of solution) with constant agitation for 1 hour.The lenses were rinsed twice with water at ambient temperature for 30minutes. The lenses were packaging in borate buffer packing solutionwith 50 ppm methyl cellulose and autoclaved at 121° C. for about 19minutes.

The lenses were worn for 8-12 hours by 7-8 human patients. No discomfortwas reported, and no adverse events or clinically significant cornealstaining or deposition was noted. Wettability was uniform at dispensing.Mild to trace non-uniform wettability was noted in some patients at thefollow-up visit.

All worn lenses were collected. The residual levels of ALA-EE wereextracted using methylene chloride and quantified via gaschromatography. The lenses contacted with 0.05% ALA-EE had an average of27 ug released and the lenses contacted with the 0.1% ALA-EE had anaverage of 51 ug released over the wear time, respectively. Thiscorresponded to a rate of release of 3.7 and 6.4 ug/hour, assuminglinear release rate.

Example 13

Using animal data from a dry eye mouse model for rewetting dropscontaining a fatty acid, it is estimated that a 4 drops per day dosageof a 0.2% ALA rewetting drop would equate to cumulated dose of 40micrograms applied to the ocular surface, taking into account theassumptions that a drop represents 50 microliters and that 90% of thedrop is lost and only 10% is retained initially on the eye (due toinitial loss upon drop application/initial drainage).

It is believed that the ester or amides of anti-inflammatory lipidmediator disclosed in the present application, have similar deliveryconcentrations to the ocular environment as the fatty acids. Thus, asimilar range of amount of ALA-EE made available to the ocular surfaceupon use of a contact lens loaded with 90 micrograms ALA-EE. Consideringhigher retention time on the eye during the use of the lens, suchamounts of ALA-EE released may thus provide better efficacy than whenusing a rewetting drop.

Reference throughout this specification to “one embodiment,” “certainembodiments,” “one or more embodiments,” “further embodiment,” or “anembodiment” means that a particular feature, structure, material, orcharacteristic described in connection with the embodiment is includedin at least one embodiment of the invention. Thus, the appearances ofthe phrases such as “in one or more embodiments,” “in certainembodiments,” “in one embodiment” or “in an embodiment” in variousplaces throughout this specification are not necessarily referring tothe same embodiment of the invention. Furthermore, the particularfeatures, structures, materials, or characteristics may be combined inany suitable manner in one or more embodiments.

Although the invention herein has been described with reference toparticular embodiments, it is to be understood that these embodimentsare merely illustrative of the principles and applications of thepresent invention. It will be apparent to those skilled in the art thatvarious modifications and variations can be made to the method andapparatus of the present invention without departing from the spirit andscope of the invention. Thus, it is intended that the present inventioninclude modifications and variations that are within the scope of theappended claims and their equivalents.

What is claimed is:
 1. An ophthalmic device for the treatment of ocularconditions comprising: a hydrogel contact lens incorporating an ester ofan anti-inflammatory lipid mediator that is a reaction product of theanti-inflammatory lipid mediator and a monohydric alcohol selected fromthe group consisting of eicosapentaenoic acid ethyl ester andalpha-linolenic acid ethyl ester and mixtures thereof; wherein theophthalmic device is contained in a sealed package in which the contactlens incorporating the ester of the anti-inflammatory lipid mediator isstored in the presence of a packing solution.
 2. The ophthalmic deviceof claim 1, wherein the ester or amide is present in the contact lens ina therapeutically effective amount.
 3. The ophthalmic device of claim 1,wherein the ester is present in the contact lens in an amount in therange of about 0.01% to 0.5% by weight.
 4. The ophthalmic device ofclaim 1, wherein the packing solution is water-based.
 5. The ophthalmicdevice of claim 4, wherein the packing solution further comprises one ormore of the following: methyl cellulose, a surfactant, a wetting agent,a chelant, and an antioxidant.
 6. The ophthalmic device of claim 5,wherein the packing solution comprises one or more ingredients selectedfrom the group consisting of methyl cellulose, Tyloxapol™, methylgluceth-20, Vitamin E, diethylenetriaminepentaacetic acid, boric acid,sodium borate, and sodium chloride.
 7. The ophthalmic device of claim 1,wherein upon wearing of the contact lens on-eye, the ester releases overtime.
 8. The ophthalmic device of claim 7 comprising a release rate ofapproximately 3 micrograms per hour after a first hour of wearing. 9.The ophthalmic device of claim 1, wherein the ester is incorporated intothe contact lens in an amount in the range of about 50 to about 620micrograms per lens.
 10. The ophthalmic device of claim 1, wherein thehydrogel contact lens is selected from the group consisting ofbalafilcon, acquafilcon, lotrafilcon, comfilcon, galyfilcon, senofilcon,narafilcon, falcon II 3, or asmofilcon A.
 11. The ophthalmic device ofclaim 1 wherein the ester is eicosapentaenoic acid ethyl ester.
 12. Theophthalmic device of claim 1 wherein the ester is alpha-linolenic acidethyl ester.